Cell proliferation ability was measured at 0, 24, 48 and 72 h after transfection using Counting Kit-8 (CCK-8) (Sangon Biotech, China) according to the instruction of manufacturer. Cell apoptosis rate was measured at 48 h by FITC Annexin V Apoptosis Detection Kit II with propidium iodide (AV/PI) (BD, USA) according to the instruction of manufacturer. Apoptotic protein Cleaved Caspase 3 (C-Caspase 3) and anti-apoptotic protein B-cell lymphoma-2 (Bcl-2) were measured at 48 h using Western Blot. Cell migration rate was calculated by dividing migration distance across the gap at 24 h by that at 0 h under microscope (Olympus, Japan) using wound healing assay referring to a previous report (12 (link)). In addition, invasive cell count was calculated using Matrigel invasion assay with Matrigel basement membrane matrix (BD, USA) and 8-um Polyvinylidene Fluoride contained transwell filter chamber (Coring, USA) referring to a previous report (12 (link)).
Protocol detail
Comprehensive Cellular Characterization Protocol
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FITC Annexin V Apoptosis Detection Kit II with propidium iodide (AV/PI)
Matrigel invasion assay
Matrigel basement membrane matrix
Annexin ii
Anti apoptotic protein
Apoptotic
Assay
B cell lymphoma
Basement membrane
Caspase 3
Cell
Cell migration
Cell proliferation
Fitc
Matrigel
Microscope
Polyvinylidene fluoride
Propidium iodide
Protein
Transfection
Western blot
Corresponding Organization : First Affiliated Hospital of Anhui Medical University
Other organizations : University of Science and Technology of China
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Variable analysis
independent variables
- Transfection
- Cell proliferation ability at 0, 24, 48 and 72 h
- Cell apoptosis rate at 48 h
- Apoptotic protein Cleaved Caspase 3 (C-Caspase 3) and anti-apoptotic protein B-cell lymphoma-2 (Bcl-2) at 48 h
- Cell migration rate at 24 h
- Invasive cell count
- Counting Kit-8 (CCK-8) (Sangon Biotech, China)
- FITC Annexin V Apoptosis Detection Kit II with propidium iodide (AV/PI) (BD, USA)
- Western Blot
- Wound healing assay referring to a previous report (12 (link))
- Matrigel invasion assay with Matrigel basement membrane matrix (BD, USA) and 8-um Polyvinylidene Fluoride contained transwell filter chamber (Coring, USA) referring to a previous report (12 (link))
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